Mucor circinelloides capable of promoting root development of salvia miltiorrhiza and tanshinone synthesis and application of mucor circinelloides
A technology of Mucor crimperides and tanshinone, applied in the fields of application, microbe-based methods, chemicals for biological control, etc., can solve the problems of lack of drug sources, ecological damage, slow growth of medicinal plants, etc., and achieve large applications value, promote biosynthesis and accumulation, and promote the effect of Danshen root development
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Embodiment 1
[0052] Example 1: Isolation, cultivation, identification and preservation of Mucor circinelloides (M.circinelloides) DF20:
[0053] Endophytic fungal strain DF20 of the present invention is obtained by the following steps of isolation:
[0054] 1) Select Salvia miltiorrhiza (S. miltiorrhiza) plants of suitable size and healthy growth indoors, cut off the root of Salvia miltiorrhiza with a length of about 5 cm, and remove the soil attached to the root surface as much as possible.
[0055] 2) Rinse the intercepted root segment of Salvia miltiorrhiza with sterilized PBS solution for 2 times and then with sterile water for 2 times. Then soak the root section of Salvia miltiorrhiza in 200mL of sterilized PBS solution, and vibrate ultrasonically for 10min.
[0056] PBS solution (Sangon Biology) formula: NaCl 136.89mM; KCl 2.67mM; NaCl 2 HPO 4 8.1mM; KH 2 PO 4 1.76mM; 121°C, 15min, sterilized.
[0057] 3) Take out the root of Salvia miltiorrhiza, wash it with sterile water (50...
Embodiment 2
[0065] Embodiment 2: the acquisition of the aseptic vaccine of Salvia miltiorrhiza
[0066] The aseptic vaccine of Salvia miltiorrhiza used was obtained through the following steps:
[0067] 1) Select plump seeds of Salvia miltiorrhiza, rinse with running water, treat with 75% alcohol, rinse with sterile water, treat with 5% NaClO solution, rinse with sterile water, absorb excess water with sterile filter paper, and place in a tissue culture room at 28°C Germination culture, the medium is MS medium (pH 5.8-6.0), the culture conditions are temperature 25-28°C, light intensity 2800-3000lx, and light time 14h / d.
Embodiment 3
[0069] Embodiment 3: binary co-cultivation of endophytic fungi and sterile vaccines of Salvia miltiorrhiza
[0070] 1) Get the Mucor circinelloides (M.circinelloides) DF20 gained in Example 1, under aseptic conditions, pick the mycelium of an inoculation loop, inoculate it in 100ml sterile PDA liquid medium, place it at 28°C, 150rpm shaker culture for 3-5 days, take an appropriate amount of bacterial solution and dilute it with sterile PDA medium to form a bacterial suspension (10 6 ~10 7 CFU / mL).
[0071] 2) Take the aseptic seedlings of 3-4 leaf age Salvia miltiorrhiza obtained in Example 2, and transplant them into the aseptic culture medium (peat: vermiculite: perlite=3:1:1) equipped with 100ml MS medium, Cultivate in a tissue culture room at 25°C for 10-15 days, take 1ml of the DF20 bacterial suspension obtained in step 1) and inoculate the roots of Salvia miltiorrhiza seedlings, and inoculate 1ml of sterile PDA liquid culture medium grown on a shaker under the same con...
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